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M-PER Mammalian Protein Extraction Reagent


A mammalian cell lysis solution that eliminates scraping, freeze-thaw and sonication methods.

Thermo Scientific M-PER Mammalian Protein Extraction Reagent

Thermo Scientific M-PER Mammalian Protein Extraction Reagent is designed to provide highly efficient total soluble protein extraction from cultured mammalian cells.

The complete cell lysis reagent contains a mild, nondenaturing detergent that dissolves cell membranes and extracts the total soluble cellular protein from most cellular compartments in only 5 minutes and requires little or no additional mechanical disruption. M-PER Reagent is formulated for minimal interference with downstream biological applications. Protein extraction efficiency of M-PER Reagent has been validated using several cell lines, including primary, suspension and adherent cell types and the resulting cell lysates are compatible with many downstream assays including immunoassays, enzyme assays and a variety of common reporter assays.

Highlights:

  • Gentle – mild detergent lysis, yielding extracts that are immediately compatible with coomassie (Bradford) and BCA protein assays or SDS-PAGE
  • Compatible – extracts soluble proteins in nondenatured state, enabling direct use in immunoprecipitation and other affinity purification procedures
  • Amine-free and dialyzable – formulation ensures compatibility with subsequent assay systems
  • Convenient – lyse adherent cells directly in plate or after scraping and washing in suspension
  • Non-denaturing – maintain activity of luciferase, beta-galactosidase, CAT and other reporter genes as well or better than other suppliers' products and freeze/thaw methods

Product Details:

Cellular protein extraction is a key first step in many proteomics analysis procedures and requires cell lysis to release the proteins of interest. M-PER Reagent is a robust cell lysis method and efficiently lyses a variety of cell types and is reliable for working with precious primary cells and cells grown in suspension or adherent culture conditions. M-PER Reagent efficiency was evaluated by both protein yield and confirmation that specific target proteins from the various cellular compartments are recovered. Additionally, because the M-PER Reagent is primary amine-free and dialyzable, it is more compatible with many downstream applications than many other protein extraction methods including luciferase, beta-galactosidase and CAT assays.

Cell lysis protein yield with M-PER Reagent
Protein yield from various cell types using M-PER Mammalian Protein Extraction Reagent. Cells were harvested at 85% confluency , washed twice and collected in ice-cold PBS and counted. For each cell type, 1 x 10^6 cells were pelleted by centrifugation at 2,000 x g for 5 minutes and lysed in 1mL M-PER Reagent for 5 minutes.  The cell lysates were clarified by centrifugation at 14,000 x g for 10 minutes and the supernatant was collected and the protein concentration (µg/million cells) was determined using the Pierce BCA Protein Assay (Part No. 23227).

 

Efficient protein extraction from cellular compartments using M-PER Reagent
Protein extraction efficiency from major cellular compartments using M-PER Mammalian Protein Extraction Reagent. Lysates from established cell lines and primary cultures were prepared using M-PER Reagent and extraction efficiency from the various cellular compartments evaluated. For each target protein, 10µg of lysate was loaded for and electrophoresed by SDS PAGE, transferred to nitrocellose membrane and detected by Western blot using SuperSignal West Pico Chemiluminescent Substrate (Part No. 34080).

 

M-PER cell lysis reagent compatible with reporter assays
M-PER Reagent compatibility with reporter assays in transiently transfected mammalian cells. FM2 cells were transiently transfected with a reporter construct containing the luciferase gene. The transfected cells were lysed with either M-PER Reagent or Brand P lysis buffer and subjected to luciferase assay. For β-galactosidase and CAT assays, MDA-MB-231 cells were co-transfected with reporter constructs expressing β-Gal and CAT, respectively. The transfected cells were lysed with M-PER Reagent or the freeze-thaw method, and the lysates were assayed for activity.

References:

  1. Su, WC., et al. (2011). Rab5 and Class III phosphoinositide 3-kinase Vps34 are involved in hepatitis C virus NS4B-induced autophagy. J Virol., 85:10561-71.
  2. Landles, C., et al. (2010). Proteolysis of mutant huntingtin produces an exon 1 fragment that accumulates as an aggregated protein in neuronal nuclei in Huntington disease. J Biol Chem., 285:8808-23.
  3. Patrick, PS., et al. (2014). Dual-modality gene reporter for in vivo imaging. PNAS., 111:415-20.
  4. Gibbons, AV., et al. (2013). Intestinal GUCY2C prevents TGF-beta secretion coordinating desmoplasia and hyperproliferation in colorectal cancer. Cancer Res., 73:6654-66.
  5. Abe, S., et al. (2013). A novel targeting therapy of malignant mesothelioma using anti-podoplanin antibody. J Immunol., 190:6239-49.
  6. Eapen, A., et al. (2013). Dentin phosphophoryn activates Smad protein signaling through Ca2+-calmodulin-dependent protein kinase II in undifferentiated mesenchymal cells. J Biol Chem., 288:8585-95.
  7. Liu, G., et al. (2012). Replication fork stalling and checkpoint activation by a PKD1 locus mirror repeat polypurine-polypyrimidine (Pu-Py) tract. J Biol Chem., 287:33412-23.
  8. Li, Y., et al. (2012). Extracellular membrane-proximal domain of HAb18G/CD147 binds to metal ion-dependent adhesion site (MIDAS) motif of integrin beta1 to modulate malignant properties of hepatoma cells. J Biol Chem., 287:4759-72.
  9. Yilmaz, MB., et al. (2011). Aromatase promoter I.f is regulated by progesterone receptor in mouse hypothalamic neuronal cell lines. J Mol Endocrinol., 47:69-80.
  10. Edfeldt, K., et al. (2010). T-cell cytokines differentially control human monocyte antimicrobial responses by regulating vitamin D metabolism. PNAS, 107: 22593-8.
  11. Liao, TL., et al. (2010). Ubiquitination and deubiquitination of NP protein regulates influenza A virus RNA replication. EMBO J., 29:3879-90.
  12. Price, CTD., et al. (2010). Exploitation of conserved eukaryotic host cell farnesylation machinery by an F-box effector of Legionella pneumophila. J Exp Med., 207:1713-26.

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Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
78501 M-PER Mammalian Protein Extraction Reagent
Formulation: Proprietary detergent in 25mM bicine buffer, pH 7.6
Sufficient For: 25g of wet cells (10 million cells per 1mL of reagent)
250mL Product Instructions for product #78501 M-PER Mammalian Protein Extraction Reagent MSDS for product #78501 M-PER Mammalian Protein Extraction Reagent Certificate of Analysis for product #78501 M-PER Mammalian Protein Extraction Reagent $262.00

Add to cart
78503 M-PER Mammalian Protein Extraction Reagent
Formulation: Proprietary detergent in 25mM bicine buffer, pH 7.6
Sufficient For: 2.5g of wet cells (10 million cells per 1mL of reagent)
25mL Product Instructions for product #78503 M-PER Mammalian Protein Extraction Reagent MSDS for product #78503 M-PER Mammalian Protein Extraction Reagent Certificate of Analysis for product #78503 M-PER Mammalian Protein Extraction Reagent $67.00

Add to cart
78505 M-PER Mammalian Protein Extraction Reagent
Formulation: Proprietary detergent in 25mM bicine buffer, pH 7.6
Sufficient For: 100g of wet cells (10 million cells per 1mL of reagent)
1L Product Instructions for product #78505 M-PER Mammalian Protein Extraction Reagent MSDS for product #78505 M-PER Mammalian Protein Extraction Reagent Certificate of Analysis for product #78505 M-PER Mammalian Protein Extraction Reagent $707.00

Add to cart

Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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